Since The Intended Cooling Blood To Eliminate Mange Soup Clinical Observation And Its Mechanism Of Action Of The Treatment Of Psoriasis Vulgaris

Objective1.To observe the clinic curative effect and safety of Liangxuexiaobi Decoction(LXXB) for psoriasis vulgaris.2.To observe the cell proliferation and VEGF expression of HaCaT cells which are cultured in vitro and intervened by different concentration of Liangxuexiaobi Decoction. Discuss the effective therapy mechanism of traditional Chinese medicine in psoriasis vulgaris and provide scientific basis for asiaticoside application in clinic.MethodsⅠclinical observation1.A total of 46 psoriasis vulgaris sample patients who came from the psoriasis clinic in The NO.1 Hospital of Wuhan from August,2005 to March,2008 were observed according to the standards of psoriasis area below 30%of human's body surface area,and never accepted the therapy of traditional Chinese medicine or immunosuppressive one month before observed.2.The patients were divided into 2 groups randomly.Two groups have no obvious differrences in age,sex and PASI(P＞0.05)has comparability.The treatment group includes 26 cases which are given Liangxuexiaobi Decoction,one dose for one day,taken at two times,while the comparison group includes 20 cases which are given Yujinyinxie PIece, three times one day,with 4 pieces each time.Take 10%coal tar ointment as the local treatment both groups.We set every two weeks as a treatment course.Then the clinic curative effects of the two groups are compared according to the PASI,after observing four treatment courses consecutively.Also blood analysis,urine analysis,Hepatic function and renal function is monitored and reactions are recorded.3.Statistics:After the observation,we analyzed all the data by SPSS14.0.ⅡExperiment1.HaCaT cells were cultured in vitro in 10%DMEM culture medium.Digest HaCaT cell by 0.25%pancreas enzyme,37℃,5%CO₂,5min,after the cells were pasted with the wall of culture bottle 70%-80%.Observed the morphological change of HaCaT through the convert microscope,add appropriate culture medium to stop the digestion when the cells turn round and the cells junctions get interrupted,blow the wall of culture bottle until it becomes unicellular suspension,then inoculated to 96 hole board.2.Rats are divided into groups to make serum contained Liangxuexiaopi Decoction by intragastral.Divided 24 rats into 4 groups:blank comparison group(physiological Saline group,liangxuexiaobi decoction groups(the end concentration is 0.5g/ml,1g/ml,2g/ml),every group contain 6,half is female and the other is male.The rats of the LXXBD groups were medicated once a day,continuing for 7 days.The rats of blank compareis- on groups(physiological saline group)were medicated equal physiological saline ever- yday.(《TCM Research Design & Statistics》Hunan science and technology publicshing house 48 the method of medication about experimental animals).Collect blood 1 hour after the last intragastral,distill the serum,store at -20℃refrigerator.3.Obtain the proliferation level of the cell which is intervened by LXXBD using MTT. Inoculated cells to 96 holes board when density is 2×10⁴.Discard the old culture medium when the cells were pasted with the wall of culture bottle 70%-80%,then keep the culture medium without FBS for 4 hours,check the OD₄₉₂by the enzyme marks board after adding different concentration medicine.4.Inoculated the cells to 96 holes board when density is 1×10⁵.Discard the old culture medium when the cells were pasted with the wall of culture bottle 70%-80%,add different concentration medicine,collect supernatant after 24 hours,Check the expression of VEGF by ELISA according to the approach of Human VEGF ELISA KIT from Jingmei.Result:一.clinical observationAccording to the decline ratio of PASI,as for the treatment group,the convalesceng ate is 7.69%,the showing results rate is 57.69%,and the total showing results rate is 65.38%.While for the comparison group,the convalescing rate is 5%,showing results rate is 30%,and the total showing results rate is 35%.The treatment group total showing results rate is higher than the comparison group.The score of the treatment group is 9.64±3.86 while the score of the comparison group is 9.58±2.48 before accept treatment,we get the the score of the treatment group is 4.23±3.55 while the score of the comparison group is 5.48±3.83 at the end of the observation,there is obvious differences between the scores below(P＜0.01).二.experiment1.In the experiment of MTT,the OD₄₉₂value of blank control group is 0.961±0.028, the OD₄₉₂value of blank control group which is intervened by TGF-a is 1.163±0.073, the OD₄₉₂value of blind comparison group is 1.098±0.067,the OD492 value of different LXXBD groups sequentially are 0.908±0.041,0.914±0.096,0.553±0.159.We can see that the OD₄₉₂value of blank control group which is intervened by TGF-a is higher than the OD₄₉₂value of blank control group(P＜0.05).Following the rising of medicine concentration gradient,the cell proliferation activation gradually reduced,the cell proliferation levels of LXXBD groups are obviously lower than which is in HaCaT cell group(P＜0.05),and among LXXBD groups,there is no significance difference between the low concentration group and medium concentration group (P＞0.05).But it shows an obvious difference between high concentration group and blind comparison group or low concentration group or medium concentration group (P＜0.01).2.From the result of ELISA,we can find that the VEGF expression of blank control group is 0.473±0.142,the value of blank control group which is intervened by TGF-a is 1.001±0.261,and the value of different concentration LXXBD sequentially are 1.185±0.133,0.964±0.403,0.33±0.067.There is significant difference between blank control group which is intervened by TGF-a and blank control group(P＜0.01).the VEGF expression of LXXBD groups are obviously lower than which is in physiological saline group(P＜0.01).It shows that following the rising of medicine concentration gradient,the VEGF expression gradually reduced,and among the LXXBD groups,the values of VEGF expression also has difference(P＜0.01).By Correlation analysis,the VEGF expression is highly negatively related to medicine concentration gradient,r=-0.998,P＜0.01.It shows that LXXBD can evidently suppress the VEGF expression of HaCaT cell. Conclusion:1.The LXXBD has evident clinic curative effect for psoriasis vulgaris,it is better than the comparing group obviously.2.The serum with LXXBD could restrain the proliferation of HaCaT cell and has a good effect of reducing the VEGF secretion,bating the expression of VEGF,and controlling the angiogenesis,which might be an effective therapy mechanism for psoriasis vulgaris curing.